Human NLRP3 knockout THP-1 cell line
- Advanced Validation
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NLRP3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 58bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
AGTAVPRL, AII/AVP, Angiotensin/vasopressin receptor AII/AVP-like, C1orf7, CIAS 1, CLR1.1, Caterpiller protein 1.1, Cold autoinflammatory syndrome 1, Cold autoinflammatory syndrome 1 protein, Cryopyrin, FCAS, FCU, Familial cold autoinflammatory syndrome, LRR and PYD domains-containing protein 3, MWS, Muckle-Wells syndrome, NACHT, NACHT LRR and PYD containing protein 3, NALP 3, NALP3_HUMAN, NLR family pyrin domain containing 3, PYPAF 1, PYRIN-containing APAF1-like protein 1
- WB
Lab
Western blot - Human NLRP3 knockout THP-1 cell line (AB280063)
False colour image of Western blot : Anti-NLRP3 antibody staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab4207 was shown to bind specifically to NLRP3. A band was observed at 118 kDa in wild-type THP-1 cell lysates with no signal observed at this size in NLRP3 knockout cell line ab280063 (knockout cell lysate ab280122). To generate this image wild-type and NLRP3 knockout THP-1 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Donkey anti-Goat IgG H&L (IRDye® 800CW) preabsorbed (ab216775) and Donkey anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216778) at 1/20000 dilution.
All lanes:
Western blot - Anti-NLRP3 antibody (<a href='/en-us/products/primary-antibodies/nlrp3-antibody-ab4207'>ab4207</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 cell lysate at 20 µg
Lane 2:
NLRP3 knockout THP-1 cell lysate at 20 µg
Lane 2:
Western blot - Human NLRP3 knockout THP-1 cell line (ab280063)
Predicted band size: 118 kDa
Observed band size: 118 kDa
false
- WB
Lab
Western blot - Human NLRP3 knockout THP-1 cell line (AB280063)
False colour image of Western blot : Anti-NLRP3 antibody [EPR23094-1] staining at 1/500 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab263899 was shown to bind specifically to NLRP3. A band was observed at 118 kDa in wild-type THP-1 cell lysates with no signal observed at this size in NLRP3 knockout cell line ab280063 (knockout cell lysate ab280122). To generate this image wild-type and NLRP3 knockout THP-1 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T incubated with secondary antibodies for 1 h at room temperature washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) at 1/20000 dilution.
All lanes:
Western blot - Anti-NLRP3 antibody [EPR23094-1] (<a href='/en-us/products/primary-antibodies/nlrp3-antibody-epr23094-1-ab263899'>ab263899</a>) at 1/500 dilution
Lane 1:
Wild-type THP-1 cell lysate at 20 µg
Lane 2:
NLRP3 knockout THP-1 cell lysate at 20 µg
Lane 2:
Western blot - Human NLRP3 knockout THP-1 cell line (ab280063)
Predicted band size: 118 kDa
Observed band size: 118 kDa
false
- WB
Lab
Western blot - Human NLRP3 knockout THP-1 cell line (AB280063)
ab263899 was shown to react with NLRP3 in wild-type THP-1 cells in Western blot with loss of signal observed in a NLRP3 knockout cell line. Wild-type THP-1 and NLRP3 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab263899 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes:
Western blot - Anti-NLRP3 antibody [EPR23094-1] (<a href='/en-us/products/primary-antibodies/nlrp3-antibody-epr23094-1-ab263899'>ab263899</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 lysate at 20 µg
Lane 2:
Wild-type THP-1 lysate treated with 200 ng/ml PMA, 48 hr at 20 µg
Lane 3:
NLRP3 knock-out THP-1 lysate at 20 µg
Lane 4:
NLRP3 knock-out THP-1 lysate treated with 200 ng/ml PMA, 48 hr at 20 µg
false
- Sanger seq
Lab
Sanger Sequencing - Human NLRP3 knockout THP-1 cell line (AB280063)
58bp deletion in exon 2
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
Culture medium
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
NLRP3 is central to the body's defense mechanism by participating in forming the inflammasome complex which activates caspase-1. This activation leads to the cleavage of pro-inflammatory cytokines like IL-1β and IL-18 into their active forms. NLRP3's function is essential for mounting an effective immune response to infections and damage signals. It senses a range of stimuli including crystalline substances and cellular stress triggering an immune response. Its expression is tightly regulated to prevent excessive inflammation.
Pathways
The NLRP3 inflammasome is key in the innate immune response and inflammation pathways. It links cellular stress signals to inflammation through the activation of caspase-1 which is important for the IL-1 signaling pathway. NLRP3 interacts with other proteins like ASC (apoptosis-associated speck-like protein containing a CARD) to form the inflammasome complex which is a core part of the inflammatory signaling pathways and links innate and adaptive immunity.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB263899
Anti-NLRP3 antibody [EPR23094-1]
KO Validated|RabMAb|Recombinant|20ul selling size
![Western blot - Anti-NLRP3 antibody [EPR23094-1] (AB263899)](https://content.abcam.com/products/images/nlrp3-antibody-epr23094-1-ab263899--western-blot-img421355.jpg)
- 4
- 2
Cell Lines & Lysates
AB276116
Human CASP1 (Caspase-1) knockout THP-1 cell line
Advanced Validation
- 0
- 0
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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